A quantitative colorimetric method to evaluate the functional state of human polymorphonuclear leukocytes.
The colorimetric assay previously described by Mosmann for the measurement of cell viability and proliferation has been modified for the assessment of the functional state of human polymorphnuclear cells (PMNs). The ability of PMNs to reduce the tetrazolium salt MTT to formazan reflects directly the degree of stimulation induced by various agents. The underlying mechanism of MTT-reduction to formazan seems to be similar to that of nitroblue tetrazolium (NBT)-reduction. In contrast to the NBT-reduction assay, the formazan produced from MTT can easily be measured by an ELISA reader. Parallel experiments revealed a qualitative correlation between the concentration of formazan produced from MTT and the concentration of cytochrome C reduced by PMNs. Although oxidative burst may not be the actual lytic mechanism in cellular cytotoxicity of PMN, we also observed an association between MTT-reduction capacity and the cytotoxic activity of PMNs from normal donors in antibody dependent cellular cytotoxicity. Our results indicate that the MTT-reduction assay can be employed to estimate the functional state of polymorphnuclear granulocytes.
Oez
S
S
Laboratory of Cytokine Biology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
Platzer
E
E
Welte
K
K
D002455:Cell Division; D002470:Cell Survival; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000502:physiology; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D013778:Tetrazolium Salts; D013844:Thiazoles; D006861:Hydrogen Peroxide; C022616:thiazolyl blue; D010100:Oxygen